Figure 6.

Medium depth affects RPE lipid droplet size, number, and localization after lipid challenge. In both hypoxic (1% O₂) and high media volume (200 µL) conditions, there is an increase in the (A) number of LDs and (B) size of LDs formed after an 18-hour lipid challenge (300 µM palmitate) in hfRPE (n = 3–4 randomly selected images). (C) Immunostaining LDs (ADRP/perilipin-2, green) and nuclei (Hoechst 34580, blue) under high volume or 1% O₂ conditions. (D) The larger LDs that form under high-volume or low atmospheric O₂ conditions are predominantly basolateral (red arrowhead), adjacent to nuclei. The smaller LDs that form under normoxia are more apically localized (white arrowhead). All results from donor 6, and replication of results in an iPSC-RPE donor and another hfRPE donor is in Supplementary Figure S6. Scale bar: 10 µm.